Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Oct;70(10):6839–6846. doi: 10.1128/jvi.70.10.6839-6846.1996

Generation of recombinant adenovirus vectors with modified fibers for altering viral tropism.

V N Krasnykh 1, G V Mikheeva 1, J T Douglas 1, D T Curiel 1
PMCID: PMC190731  PMID: 8794325

Abstract

To expand the utility of recombinant adenovirus vectors for gene therapy applications, methods to alter native viral tropism to achieve cell-specific transduction would be beneficial. To this end, we are pursuing genetic methods to alter the cell recognition domain of the adenovirus fiber. To incorporate these modified fibers into mature virions, we have developed a method based on homologous DNA recombination between two plasmids. A fiber-deleted, propagation-defective rescue plasmid has been designed for recombination with a shuttle plasmid encoding a variant fiber gene. Recombination between the two plasmids results in the derivation of recombinant viruses containing the variant fiber gene. To establish the utility of this method, we constructed a recombinant adenovirus containing a fiber gene with a silent mutation. In addition, we generated an adenovirus vector containing chimeric fibers composed of the tail and shaft domains of adenovirus serotype 5 and the knob domain of serotype 3. This modification was shown to alter the receptor recognition profile of the virus containing the fiber chimera. Thus, this two-plasmid system allows for the generation of adenovirus vectors containing variant fibers. This method provides a rapid and facile means of generating fiber-modified recombinant adenoviruses. In addition, it should be possible to use this system in the development of adenovirus vectors with modified tropism to allow cell-specific targeting.

Full Text

The Full Text of this article is available as a PDF (261.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Addison C. L., Braciak T., Ralston R., Muller W. J., Gauldie J., Graham F. L. Intratumoral injection of an adenovirus expressing interleukin 2 induces regression and immunity in a murine breast cancer model. Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8522–8526. doi: 10.1073/pnas.92.18.8522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bai M., Harfe B., Freimuth P. Mutations that alter an Arg-Gly-Asp (RGD) sequence in the adenovirus type 2 penton base protein abolish its cell-rounding activity and delay virus reproduction in flat cells. J Virol. 1993 Sep;67(9):5198–5205. doi: 10.1128/jvi.67.9.5198-5205.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ballay A., Levrero M., Buendia M. A., Tiollais P., Perricaudet M. In vitro and in vivo synthesis of the hepatitis B virus surface antigen and of the receptor for polymerized human serum albumin from recombinant human adenoviruses. EMBO J. 1985 Dec 30;4(13B):3861–3865. doi: 10.1002/j.1460-2075.1985.tb04158.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berkner K. L., Schaffhausen B. S., Roberts T. M., Sharp P. A. Abundant expression of polyomavirus middle T antigen and dihydrofolate reductase in an adenovirus recombinant. J Virol. 1987 Apr;61(4):1213–1220. doi: 10.1128/jvi.61.4.1213-1220.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bett A. J., Haddara W., Prevec L., Graham F. L. An efficient and flexible system for construction of adenovirus vectors with insertions or deletions in early regions 1 and 3. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8802–8806. doi: 10.1073/pnas.91.19.8802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Boudin M. L., Boulanger P. Assembly of adenovirus penton base and fiber. Virology. 1982 Jan 30;116(2):589–604. doi: 10.1016/0042-6822(82)90151-9. [DOI] [PubMed] [Google Scholar]
  7. Bout A., Perricaudet M., Baskin G., Imler J. L., Scholte B. J., Pavirani A., Valerio D. Lung gene therapy: in vivo adenovirus-mediated gene transfer to rhesus monkey airway epithelium. Hum Gene Ther. 1994 Jan;5(1):3–10. doi: 10.1089/hum.1994.5.1-3. [DOI] [PubMed] [Google Scholar]
  8. Cardoso J. E., Branchereau S., Jeyaraj P. R., Houssin D., Danos O., Heard J. M. In situ retrovirus-mediated gene transfer into dog liver. Hum Gene Ther. 1993 Aug;4(4):411–418. doi: 10.1089/hum.1993.4.4-411. [DOI] [PubMed] [Google Scholar]
  9. Chroboczek J., Ruigrok R. W., Cusack S. Adenovirus fiber. Curr Top Microbiol Immunol. 1995;199(Pt 1):163–200. doi: 10.1007/978-3-642-79496-4_10. [DOI] [PubMed] [Google Scholar]
  10. Chu T. H., Dornburg R. Retroviral vector particles displaying the antigen-binding site of an antibody enable cell-type-specific gene transfer. J Virol. 1995 Apr;69(4):2659–2663. doi: 10.1128/jvi.69.4.2659-2663.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chu T. H., Martinez I., Sheay W. C., Dornburg R. Cell targeting with retroviral vector particles containing antibody-envelope fusion proteins. Gene Ther. 1994 Sep;1(5):292–299. [PubMed] [Google Scholar]
  12. Clayman G. L., el-Naggar A. K., Roth J. A., Zhang W. W., Goepfert H., Taylor D. L., Liu T. J. In vivo molecular therapy with p53 adenovirus for microscopic residual head and neck squamous carcinoma. Cancer Res. 1995 Jan 1;55(1):1–6. [PubMed] [Google Scholar]
  13. Cosset F. L., Morling F. J., Takeuchi Y., Weiss R. A., Collins M. K., Russell S. J. Retroviral retargeting by envelopes expressing an N-terminal binding domain. J Virol. 1995 Oct;69(10):6314–6322. doi: 10.1128/jvi.69.10.6314-6322.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Crystal R. G., McElvaney N. G., Rosenfeld M. A., Chu C. S., Mastrangeli A., Hay J. G., Brody S. L., Jaffe H. A., Eissa N. T., Danel C. Administration of an adenovirus containing the human CFTR cDNA to the respiratory tract of individuals with cystic fibrosis. Nat Genet. 1994 Sep;8(1):42–51. doi: 10.1038/ng0994-42. [DOI] [PubMed] [Google Scholar]
  15. Csete M. E., Afra R., Mullen Y., Drazan K. E., Benhamou P. Y., Shaked A. Adenoviral-mediated gene transfer to pancreatic islets does not alter islet function. Transplant Proc. 1994 Apr;26(2):756–757. [PubMed] [Google Scholar]
  16. Davidson D., Hassell J. A. Overproduction of polyomavirus middle T antigen in mammalian cells through the use of an adenovirus vector. J Virol. 1987 Apr;61(4):1226–1239. doi: 10.1128/jvi.61.4.1226-1239.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. DeMatteo R. P., Raper S. E., Ahn M., Fisher K. J., Burke C., Radu A., Widera G., Claytor B. R., Barker C. F., Markmann J. F. Gene transfer to the thymus. A means of abrogating the immune response to recombinant adenovirus. Ann Surg. 1995 Sep;222(3):229–242. doi: 10.1097/00000658-199509000-00002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Defer C., Belin M. T., Caillet-Boudin M. L., Boulanger P. Human adenovirus-host cell interactions: comparative study with members of subgroups B and C. J Virol. 1990 Aug;64(8):3661–3673. doi: 10.1128/jvi.64.8.3661-3673.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Dong J., Roth M. G., Hunter E. A chimeric avian retrovirus containing the influenza virus hemagglutinin gene has an expanded host range. J Virol. 1992 Dec;66(12):7374–7382. doi: 10.1128/jvi.66.12.7374-7382.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Emi N., Friedmann T., Yee J. K. Pseudotype formation of murine leukemia virus with the G protein of vesicular stomatitis virus. J Virol. 1991 Mar;65(3):1202–1207. doi: 10.1128/jvi.65.3.1202-1207.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Falgout B., Ketner G. Characterization of adenovirus particles made by deletion mutants lacking the fiber gene. J Virol. 1988 Feb;62(2):622–625. doi: 10.1128/jvi.62.2.622-625.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Fujiwara T., Grimm E. A., Mukhopadhyay T., Zhang W. W., Owen-Schaub L. B., Roth J. A. Induction of chemosensitivity in human lung cancer cells in vivo by adenovirus-mediated transfer of the wild-type p53 gene. Cancer Res. 1994 May 1;54(9):2287–2291. [PubMed] [Google Scholar]
  23. Ghosh-Choudhury G., Haj-Ahmad Y., Graham F. L. Protein IX, a minor component of the human adenovirus capsid, is essential for the packaging of full length genomes. EMBO J. 1987 Jun;6(6):1733–1739. doi: 10.1002/j.1460-2075.1987.tb02425.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  25. Greber U. F., Willetts M., Webster P., Helenius A. Stepwise dismantling of adenovirus 2 during entry into cells. Cell. 1993 Nov 5;75(3):477–486. doi: 10.1016/0092-8674(93)90382-z. [DOI] [PubMed] [Google Scholar]
  26. Han X., Kasahara N., Kan Y. W. Ligand-directed retroviral targeting of human breast cancer cells. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9747–9751. doi: 10.1073/pnas.92.21.9747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Henry L. J., Xia D., Wilke M. E., Deisenhofer J., Gerard R. D. Characterization of the knob domain of the adenovirus type 5 fiber protein expressed in Escherichia coli. J Virol. 1994 Aug;68(8):5239–5246. doi: 10.1128/jvi.68.8.5239-5246.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Huard J., Lochmüller H., Acsadi G., Jani A., Massie B., Karpati G. The route of administration is a major determinant of the transduction efficiency of rat tissues by adenoviral recombinants. Gene Ther. 1995 Mar;2(2):107–115. [PubMed] [Google Scholar]
  29. Jaffe H. A., Danel C., Longenecker G., Metzger M., Setoguchi Y., Rosenfeld M. A., Gant T. W., Thorgeirsson S. S., Stratford-Perricaudet L. D., Perricaudet M. Adenovirus-mediated in vivo gene transfer and expression in normal rat liver. Nat Genet. 1992 Aug;1(5):372–378. doi: 10.1038/ng0892-372. [DOI] [PubMed] [Google Scholar]
  30. Jolly D. Viral vector systems for gene therapy. Cancer Gene Ther. 1994 Mar;1(1):51–64. [PubMed] [Google Scholar]
  31. Kasahara N., Dozy A. M., Kan Y. W. Tissue-specific targeting of retroviral vectors through ligand-receptor interactions. Science. 1994 Nov 25;266(5189):1373–1376. doi: 10.1126/science.7973726. [DOI] [PubMed] [Google Scholar]
  32. Le Gal La Salle G., Robert J. J., Berrard S., Ridoux V., Stratford-Perricaudet L. D., Perricaudet M., Mallet J. An adenovirus vector for gene transfer into neurons and glia in the brain. Science. 1993 Feb 12;259(5097):988–990. doi: 10.1126/science.8382374. [DOI] [PubMed] [Google Scholar]
  33. Liu T. J., Zhang W. W., Taylor D. L., Roth J. A., Goepfert H., Clayman G. L. Growth suppression of human head and neck cancer cells by the introduction of a wild-type p53 gene via a recombinant adenovirus. Cancer Res. 1994 Jul 15;54(14):3662–3667. [PubMed] [Google Scholar]
  34. Louis N., Fender P., Barge A., Kitts P., Chroboczek J. Cell-binding domain of adenovirus serotype 2 fiber. J Virol. 1994 Jun;68(6):4104–4106. doi: 10.1128/jvi.68.6.4104-4106.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Maeda H., Danel C., Crystal R. G. Adenovirus-mediated transfer of human lipase complementary DNA to the gallbladder. Gastroenterology. 1994 Jun;106(6):1638–1644. doi: 10.1016/0016-5085(94)90421-9. [DOI] [PubMed] [Google Scholar]
  36. Mastrangeli A., O'Connell B., Aladib W., Fox P. C., Baum B. J., Crystal R. G. Direct in vivo adenovirus-mediated gene transfer to salivary glands. Am J Physiol. 1994 Jun;266(6 Pt 1):G1146–G1155. doi: 10.1152/ajpgi.1994.266.6.G1146. [DOI] [PubMed] [Google Scholar]
  37. Mathias P., Wickham T., Moore M., Nemerow G. Multiple adenovirus serotypes use alpha v integrins for infection. J Virol. 1994 Oct;68(10):6811–6814. doi: 10.1128/jvi.68.10.6811-6814.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. McGrory W. J., Bautista D. S., Graham F. L. A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5. Virology. 1988 Apr;163(2):614–617. doi: 10.1016/0042-6822(88)90302-9. [DOI] [PubMed] [Google Scholar]
  39. Michael S. I., Hong J. S., Curiel D. T., Engler J. A. Addition of a short peptide ligand to the adenovirus fiber protein. Gene Ther. 1995 Nov;2(9):660–668. [PubMed] [Google Scholar]
  40. Michael S. I., Huang C. H., Rømer M. U., Wagner E., Hu P. C., Curiel D. T. Binding-incompetent adenovirus facilitates molecular conjugate-mediated gene transfer by the receptor-mediated endocytosis pathway. J Biol Chem. 1993 Apr 5;268(10):6866–6869. [PubMed] [Google Scholar]
  41. Mitani K., Graham F. L., Caskey C. T. Transduction of human bone marrow by adenoviral vector. Hum Gene Ther. 1994 Aug;5(8):941–948. doi: 10.1089/hum.1994.5.8-941. [DOI] [PubMed] [Google Scholar]
  42. Mittal S. K., McDermott M. R., Johnson D. C., Prevec L., Graham F. L. Monitoring foreign gene expression by a human adenovirus-based vector using the firefly luciferase gene as a reporter. Virus Res. 1993 Apr;28(1):67–90. doi: 10.1016/0168-1702(93)90090-a. [DOI] [PubMed] [Google Scholar]
  43. Moullier P., Friedlander G., Calise D., Ronco P., Perricaudet M., Ferry N. Adenoviral-mediated gene transfer to renal tubular cells in vivo. Kidney Int. 1994 Apr;45(4):1220–1225. doi: 10.1038/ki.1994.162. [DOI] [PubMed] [Google Scholar]
  44. Novelli A., Boulanger P. A. Assembly of adenovirus type 2 fiber synthesized in cell-free translation system. J Biol Chem. 1991 May 15;266(14):9299–9303. [PubMed] [Google Scholar]
  45. Novelli A., Boulanger P. A. Deletion analysis of functional domains in baculovirus-expressed adenovirus type 2 fiber. Virology. 1991 Nov;185(1):365–376. doi: 10.1016/0042-6822(91)90784-9. [DOI] [PubMed] [Google Scholar]
  46. Philipson L., Lonberg-Holm K., Pettersson U. Virus-receptor interaction in an adenovirus system. J Virol. 1968 Oct;2(10):1064–1075. doi: 10.1128/jvi.2.10.1064-1075.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Russell S. J., Hawkins R. E., Winter G. Retroviral vectors displaying functional antibody fragments. Nucleic Acids Res. 1993 Mar 11;21(5):1081–1085. doi: 10.1093/nar/21.5.1081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Siegfried W. Perspectives in gene therapy with recombinant adenoviruses. Exp Clin Endocrinol. 1993;101(1):7–11. doi: 10.1055/s-0029-1211201. [DOI] [PubMed] [Google Scholar]
  49. Smythe W. R., Kaiser L. R., Hwang H. C., Amin K. M., Pilewski J. M., Eck S. J., Wilson J. M., Albelda S. M. Successful adenovirus-mediated gene transfer in an in vivo model of human malignant mesothelioma. Ann Thorac Surg. 1994 Jun;57(6):1395–1401. doi: 10.1016/0003-4975(94)90090-6. [DOI] [PubMed] [Google Scholar]
  50. Somia N. V., Zoppé M., Verma I. M. Generation of targeted retroviral vectors by using single-chain variable fragment: an approach to in vivo gene delivery. Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7570–7574. doi: 10.1073/pnas.92.16.7570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Stevenson S. C., Rollence M., White B., Weaver L., McClelland A. Human adenovirus serotypes 3 and 5 bind to two different cellular receptors via the fiber head domain. J Virol. 1995 May;69(5):2850–2857. doi: 10.1128/jvi.69.5.2850-2857.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Stratford-Perricaudet L. D., Makeh I., Perricaudet M., Briand P. Widespread long-term gene transfer to mouse skeletal muscles and heart. J Clin Invest. 1992 Aug;90(2):626–630. doi: 10.1172/JCI115902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Takeuchi Y., Cosset F. L., Lachmann P. J., Okada H., Weiss R. A., Collins M. K. Type C retrovirus inactivation by human complement is determined by both the viral genome and the producer cell. J Virol. 1994 Dec;68(12):8001–8007. doi: 10.1128/jvi.68.12.8001-8007.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Trapnell B. C., Gorziglia M. Gene therapy using adenoviral vectors. Curr Opin Biotechnol. 1994 Dec;5(6):617–625. doi: 10.1016/0958-1669(94)90084-1. [DOI] [PubMed] [Google Scholar]
  55. Valsesia-Wittmann S., Drynda A., Deléage G., Aumailley M., Heard J. M., Danos O., Verdier G., Cosset F. L. Modifications in the binding domain of avian retrovirus envelope protein to redirect the host range of retroviral vectors. J Virol. 1994 Jul;68(7):4609–4619. doi: 10.1128/jvi.68.7.4609-4619.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Weiss R. A., Boettiger D., Murphy H. M. Pseudotypes of avian sarcoma viruses with the envelope properties of vesicular stomatitis virus. Virology. 1977 Feb;76(2):808–825. doi: 10.1016/0042-6822(77)90261-6. [DOI] [PubMed] [Google Scholar]
  57. Wickham T. J., Carrion M. E., Kovesdi I. Targeting of adenovirus penton base to new receptors through replacement of its RGD motif with other receptor-specific peptide motifs. Gene Ther. 1995 Dec;2(10):750–756. [PubMed] [Google Scholar]
  58. Wickham T. J., Filardo E. J., Cheresh D. A., Nemerow G. R. Integrin alpha v beta 5 selectively promotes adenovirus mediated cell membrane permeabilization. J Cell Biol. 1994 Oct;127(1):257–264. doi: 10.1083/jcb.127.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Wickham T. J., Mathias P., Cheresh D. A., Nemerow G. R. Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell. 1993 Apr 23;73(2):309–319. doi: 10.1016/0092-8674(93)90231-e. [DOI] [PubMed] [Google Scholar]
  60. Yee J. K., Miyanohara A., LaPorte P., Bouic K., Burns J. C., Friedmann T. A general method for the generation of high-titer, pantropic retroviral vectors: highly efficient infection of primary hepatocytes. Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9564–9568. doi: 10.1073/pnas.91.20.9564. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES